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PD L1 Monoclonal Antibody (PDL1)

Purified Mouse Monoclonal Antibody (Mab)

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All lanes : Anti-PDL-1 Antibody at 0.5-1µg/ml dilution Lane 1: 293T/PDL1 whole cell lysate Lane 2: 293T whole cell lysate Lane 3: NCI-H1975 whole cell lysate Lane 4: MDA-MB231 whole cell lysate Lane 5: MCF-7 whole cell lysate Lane 6: NCI-H226 whole cell lysate Lysates/proteins at 30 µg per lane. Secondary Goat Anti-Mouse IgG, (H+L),Peroxidase conjµgated at 1/5000 dilution. Predicted band size : 32 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemical analysis of PDL-1 in human non-small cell lung carcinoma sections(IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) by abgent test. Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Immunohistochemical analysis of PDL-1 in human non-small cell lung carcinoma sections(IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) by Dako test. Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Immunohistochemical analysis of PDL-1 in human non-small cell lung carcinoma sections(IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) by Leica test. Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Immunohistochemical analysis of PDL-1 in human tonsil tissue sections(IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) by abgent test. Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Immunohistochemical analysis of PDL-1 in human tonsil tissue sections(IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) by Dako test. Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Immunohistochemical analysis of PDL-1 in MCF-7 cell (left) and NCI-H226（right） cell sections by abgent test . Cell was fixed with formaldehyde and blocked with super block for 10 minutes at room temperature; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Immunohistochemical analysis of PDL-1 in untransfected(left) or transfected(right) with 293T cell sections by abgent test . Cell was fixed with formaldehyde; antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (0.85µg/ml) for 1 hours at room temperature. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
AW5698 staining PD-L1 in human lung squamous carcinoma sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
Overlay histogram showing isotype control(blue line), 293T-PDL1(green line), MDA-MB-231(red line), NCI-H226(black line) cells stained with PD L1 Antibody. The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NH174309) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was Mouse IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Immunohistochemical analysis of paraffin-embedded Human tonsil section using PDL1(Cat#AW5698). AW5698 was diluted at 1:500 dilution. A undiluted biotinylated goat polyvalent antibody was used as the secondary, followed by DAB staining.

SPECIFICATION
CITATIONS: 3
PROTOCOLS
BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immuno electron microscopy EIA=Enzyme Immunoassay	WB, IHC-P, FC, IHC-P-Leica
Primary Accession	Q9NZQ7
Reactivity	Human
Predicted	Human
Host	Mouse
Clonality	monoclonal
Calculated MW	33275 Da
Isotype	IgG1
Antigen Source	HUMAN

Additional Information
Gene ID	29126
Other Names	Programmed cell death 1 ligand 1, PD-L1, PDCD1 ligand 1, Programmed death ligand 1, B7 homolog 1, B7-H1, CD274, CD274, B7H1, PDCD1L1, PDCD1LG1, PDL1, PDL-1
Dilution	WB~~1:500-1:1000 IHC-P~~1:100 IHC-P-Leica~~1:100-1:600 FC~~1:25 IHC~~1:500
Target/Specificity	This PD L1 antibody is generated from a mouse immunized with a KLH conjugated synthetic peptide between 256-290 amino acids from the human region of human PD L1.
Format	Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein G column, followed by dialysis against PBS.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	PD L1 Monoclonal Antibody (PDL1) is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	CD274 (HGNC:17635)
Function	Plays a critical role in induction and maintenance of immune tolerance to self (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:31399419). As a ligand for the inhibitory receptor PDCD1/PD-1, modulates the activation threshold of T-cells and limits T-cell effector response (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:36727298). Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (PubMed:10581077). Can also act as a transcription coactivator: in response to hypoxia, translocates into the nucleus via its interaction with phosphorylated STAT3 and promotes transcription of GSDMC, leading to pyroptosis (PubMed:32929201).
Cellular Location	Cell membrane; Single-pass type I membrane protein. Early endosome membrane; Single-pass type I membrane protein. Recycling endosome membrane; Single-pass type I membrane protein. Nucleus. Note=Associates with CMTM6 at recycling endosomes, where it is protected from being targeted for lysosomal degradation (PubMed:28813417). Translocates to the nucleus in response to hypoxia via its interaction with phosphorylated STAT3 (PubMed:32929201). [Isoform 2]: Endomembrane system; Single-pass type I membrane protein
Tissue Location	Highly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes.

Research Areas

Citations ( 0 )

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

Background

Involved in the costimulatory signal, essential for T- cell proliferation and production of IL10 and IFNG, in an IL2- dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production.

References

Dong H.,et al.Nat. Med. 5:1365-1369(1999).
Freeman G.J.,et al.J. Exp. Med. 192:1027-1034(2000).
He X.-H.,et al.Acta Pharmacol. Sin. 26:462-468(2005).
Chi X.-Y.,et al.Submitted (NOV-2005) to the EMBL/GenBank/DDBJ databases.
Ota T.,et al.Nat. Genet. 36:40-45(2004).

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$ 380.00

Cat# AW5698-U100

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